The present invention relates to novel chemical compounds, to a process for their preparation and to their use as medicaments, in particular for the prevention and/or therapy of ischaemia and reperfusion damage.
The elucidation of the molecular mechanism of cell death is the subject of intense biomedical research efforts. The aim is to find specifically active compounds which have modulating action in this process. When the individual biochemical steps resulting in cell death were examined, attention was drawn to poly(ADP-ribose)-synthetase (PARS), a protein which is expressed strongly in the cell nucleus and which is involved in deoxyribonucleic acid (DNA) damage repair [Szabo and Dawson, Trends in Pharmacological Sciences, 19, 287-298 (1998)].
Activation of PARS plays an important role in N-methyl-D-aspartate (NMDA)- and NO-induced neurotoxicity [Zhang et al., Science, 263, 687-689 (1994); Wallis et al., NeuroReport, 5, 245-248 (1993)], cerebral ischaemia [Endres et al., J. Cereb. Blood Flow Metabol., 17, 1143-1151 (1997)], traumatic brain injuries [Wallis et al., Brain Res., 710, 169-177 (1996)] and ischaemia/reperfusion damage to heart and skeletal muscle [Thiemermann et al., Proc. Nat. Acad. Sci., 94, 679-683 (1997)]. In addition, inhibition of PARS appears to have a positive effect on the therapy of arthritis [Szaboet al., Japanese J. Pharm., 75, Supp. I:102 (1997)], diabetes [Shimabukuro et al., J. Clin. Invest., 100, 290-295 (1997)] and endotoxic or septic shock [Zingarelli et al., Shock, 5, 258-264 (1996)], radiosensitization of hypoxic tumour cells [Weltin et al., Oncol. Res., 6, 399-403 (1994)], chronic colitis [Jijon et al., Am. J. Physiol. Gastrointest. Liver Physiol., 279, G641-51 (2000)], sudden deafness [Tabuchi et al., Ann. Otol. Rhinol. Laryngol., 110(2), 118-21 (2001)], inflammatory pulmonary disorders, such as, for example, asthma and chronic bronchitis [Cuzzocrea et al., Eur. J. Pharm., 342, 67-76 (1998)] and cancer.
PARS, an enzyme which constructs polymeric ADP-ribose units from nicotinamide adenosine dinucleotide (NAD+) as substrate, is activated when the DNA is damaged by single- or double-strand breaks. The polymeric ADP-ribose units formed are attached both to PARS itself and to other proteins, for example histones, topoisomerases and polymerases.
Increased activation of PARS results in a massive NAD+ consumption. The marked decrease of the NAD+ concentration and the resulting impediment of ATP synthesis (decrease of the ATP concentration) causes deterioration of the energetic state of the cell, which may lead to premature cell death (necrosis).
In the heart, reperfusion of ischaemic myocardium results in the generation of radicals, neutrophil infiltration, destruction of the myocardial tissue structure, contraction dysfunctions and necrosis. The H2O2 generated during the reperfusion phase reacts rapidly with NO, forming peroxynitrite. NO, peroxynitrite and H2O2 cause DNA strand breaks, thus resulting in overstimulation of PARS.
A further important point in the case of reperfusion damage is the accumulation of neutrophils in the reperfused myocardium. Activation of PARS increases the infiltration of neutrophils by stimulating the expression of P-selectin and ICAM-1.
Healthy PARS knock-out mice capable of reproduction are substantially protected against reperfusion damage. Infiltration of neutrophils is reduced by 50% and the structure of the myocardial tissue remains intact during the reperfusion phase.
In cases of ischaemia and reperfusion damage to heart and brain, low-molecular-weight PARS inhibitors, such as, for example, 3-aminobenzamide and 1,5-dihydroxyisoquinoline, protect the tissue against necrotic cell death (reduction of the infarct size by 30 to 48%) and delay myocardial and neuronal dysfunction.
However, the PARS inhibitors hitherto tested in animal experiments have various disadvantages. Thus, for example, 3-aminobenzamide is an unspecific PARS inhibitor which also inhibits cytochrome P450 (Eriksson et al., Toxicology and applied Pharmacology, 136, 324-331 (1996)); in contrast, 5-iodo-6-amino-1,2-benzopyrone has serious side-effects (Szabo and Dawson, Trends in Pharmacol. Sciences, 19, 287-298 (1998)). Moreover, most inhibitors are not very potent and are therefore only efficacious in animals at a relatively high dosage (Thiemermann et al., Proc. Natl. Acad. Sci., 94, 679-683 (1997)).
JP-A-032645679 and Chem. Pharm. Bull. 38 (10), 2726-2732 (1990) disclose bicyclic 2,4-(1H,3H)-pyrimidinediones as 5-HT2 antagonists for the treatment of cardiovascular diseases, depression and other mental disorders. U.S. Pat. No. 5,859,014 discloses tetrahydroquinazolinedione derivatives as xcex11 adrenergic receptor antagonists for the treatment of prostate hypertrophy. WO-A-00/42025 describes dihydropyrimidinones as PARS inhibitors. DE-A-1959705 and DE-A-2126148 list uracil derivatives for preparing crop protection agents. DE-A-2142317 mentions uracil derivatives having hypnotic properties. Furthermore, various bridged uracils are described in the literature as nucleoside analogues with potential antiviral action (for example Nucleosides Nucleotides 13 (1-3), 177-196; 13 (4), 891-902 (1994) and J. Med. Chem. 39 (3), 789-795 (1996)).
Accordingly, it is an object of the present invention to provide novel substances for the prevention and/or therapy of disorders, in particular of ischaemia and reperfusion damage.
Here, the compounds according to the invention presumably act as inhibitors of poly(ADP-ribose)-synthetase (PARS).
The present invention relates to compounds of the formula (I) 
in which
A represents a ring member selected from the group consisting of:
xe2x80x94Dxe2x80x94,
xe2x80x94CH2xe2x80x94Dxe2x80x94,
xe2x80x94Dxe2x80x94CH2xe2x80x94,
xe2x80x94CHxe2x95x90CHxe2x80x94CH2xe2x80x94,
xe2x80x94CH2xe2x80x94CHxe2x95x90CHxe2x80x94,
xe2x80x94CH2xe2x80x94CH2xe2x80x94Dxe2x80x94,
xe2x80x94Dxe2x80x94CH2xe2x80x94CH2 and
xe2x80x94CH2xe2x80x94Dxe2x80x94CH2xe2x80x94, in which
D represents xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94,
X represents (C2-C10)-alkylene or (C3-C8)-cycloalkylene which are optionally mono- or polysubstituted, independently of one another, by substituents selected from the group consisting of (C1-C6)-alkoxy, hydroxyl, amino, mono- and di-(C1-C6)-alkylamino and oxo,
R1 represents hydrogen, (C1-C6)-alkyl which is optionally mono- or polysubstituted by halogen, or represents (C3-C8)-cycloalkyl,
R2 represents a group of the formula xe2x80x94SO2xe2x80x94R4, xe2x80x94SO2xe2x80x94NR5R6, xe2x80x94COxe2x80x94R7, xe2x80x94COxe2x80x94NR8R9 or xe2x80x94COxe2x80x94OR10, in which
R4 represents (C1-C6)-alkyl or (C3-C8)-cycloalkyl which are optionally substituted by (C1-C4)-alkoxy, (C6-C10)-aryl, 5- to 10-membered heteroaryl having up to 4 heteroatoms from the group consisting of N, O and/or S, or up to trisubstituted by halogen, where the aryl or heteroaryl radicals for their part are optionally substituted by hydroxyl, (C1-C4)-alkoxy, (C1-C4)-alkyl, halogen, cyano or nitro, or represents a group of the formula
xe2x80x94Gxe2x80x94E
xe2x80x83in which
E represents (C6-C10)-aryl or a 5- to 13-membered saturated, partially unsaturated or aromatic heterocycle having up to four heteroatoms from the group consisting of N, O and S, where the ring systems are in each case optionally substituted up to five times by identical or different substituents selected from the group consisting of nitro, cyano, halogen, optionally benzamido-substituted (C1-C6)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, oxo, (C1-C6)-alkoxy, carboxyl, (C1-C6)-alkoxycarbonyl, (C1-C6)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C6)-alkylaminocarbonyl, (C1-C6)-alkanoylamido, (C1-C6)-alkylsulphonyl, (C1-C6)-alkylthio, optionally (C1-C4)-alkyl-, halogen- or nitro-substituted phenylsulphonyl, 
xe2x80x83and
G is absent or represents (C6-C10)-arylene or 5- to 10-membered heteroarylene having up to 4 heteroatoms from the group consisting of N, O and S, which are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, halogen, (C1-C6)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, (C1-C6)-alkoxy, carboxyl, (C1-C6)-alkoxycarbonyl, (C1-C6)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C6)-alkylaminocarbonyl, (C1-C6)-alkanoylamido, (C1-C6)-alkylsulphonyl and (C1-C6)-alkylthio,
R5 and R6 independently of one another each represent hydrogen, (C3-C8)-cycloalkyl, (C1-C6)-alkyl, (C6-C10)-aryl or 5- to 10-membered heteroaryl which, independently of one another, are in each case optionally substituted by (C1-C4)-alkoxy, hydroxyl, cyano, carboxyl, (C1-C4)-alkoxycarbonyl, up to three times by halogen, by (C6-C10)-aryl or by 5- to 10-membered heteroaryl, where the aryl and hetaryl radicals for their part are optionally substituted by hydroxyl, (C1-C4)-alkoxy, (C1-C4)-alkyl, halogen, cyano or nitro, or
R5 and R6 together with the nitrogen atom to which they are attached form a saturated 3- to 7-membered heterocycle in which optionally one carbon ring member is replaced by a further heteroatom from the group consisting of nitrogen, oxygen and sulphur and which is optionally substituted by (C1-C4)-alkyl, hydroxyl, (C1-C4)-alkoxy, oxo, carboxyl or (C1-C4)-alkoxycarbonyl,
R7 represents (C1-C6)-alkyl or (C3-C8)-cycloalkyl which are optionally substituted by (C1-C4)-alkoxy, (C6-C10)-aryl, 5- to 10-membered heteroaryl having up to 4 heteroatoms from the group consisting of N, O and S or up to three times by halogen, where the aryl or heteroaryl radicals for their part are optionally substituted by hydroxyl, (C1-C4)-alkoxy, (C1-C4)-alkyl, halogen, cyano or nitro, or represents a group of the formula
xe2x80x94Gxe2x80x94E
xe2x80x83in which
E represents (C6-C10)-aryl or a 5- to 13-membered saturated, partially unsaturated or aromatic heterocycle having up to four heteroatoms from the group consisting of N, O and S, where the ring systems are in each case optionally substituted up to five times by identical or different substituents selected from the group consisting of nitro, cyano, halogen, optionally benzamido-substituted (C1-C6)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, oxo, (C1-C6)-alkoxy, carboxyl, (C1-C6)-alkoxycarbonyl, (C1-C6)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C6)-alkylaminocarbonyl, (C1-C6)-alkanoylamido, (C1-C6)-alkylsulphonyl, (C1-C6)-alkylthio and optionally (C1-C4)-alkyl-, halogen- or nitro-substituted phenylsulphonyl, and
G is absent or represents (C6-C10)-arylene or 5- to 10-membered heteroarylene having up to 4 heteroatoms from the group consisting of N, O and S, which are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, halogen, (C1-C6)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, (C1-C6)-alkoxy, carboxyl, (C1-C6)-alkoxycarbonyl, (C1-C6)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C6)-alkylaminocarbonyl, (C1-C6)-alkanoylamido, (C1-C6)-alkylsulphonyl and (C1-C6)-alkylthio,
R8 and R9 independently of one another each represent hydrogen, (C3-C8)-cycloalkyl or (C1-C6)-alkyl which can in each case optionally be substituted by (C1-C4)-alkoxy, hydroxyl, cyano, carboxyl, (C1-C4)-alkoxycarbonyl, up to three times by halogen, by (C6-C10)-aryl or by 5- to 10-membered heteroaryl, where the aryl and hetaryl radicals for their part are optionally substituted by hydroxyl, (C1-C4)-alkoxy, (C1-C4)-alkyl, halogen, cyano or nitro, or
R8 and R9 together with the nitrogen atom to which they are attached form a saturated 3- to 7-membered heterocycle in which optionally one carbon ring member is replaced by a further heteroatom from the group consisting of nitrogen, oxygen and sulphur and which is optionally substituted by (C1-C4)-alkyl, hydroxyl, (C1-C4)-alkoxy, oxo, carboxyl or (C1-C4)-alkoxycarbonyl,
R10 represents (C1-C6)-alkyl, (C3-C8)-cycloalkyl or (C6-C10)-aryl which are optionally mono- or polysubstituted by hydroxyl, (C1-C4)-alkoxy, halogen, cyano or nitro, or
R1 and R2 together with the nitrogen atom to which they are attached form a saturated 5- to 10-membered mono- or bicyclic heterocycle, which has a carbonyl or sulphonyl group directly adjacent to the nitrogen atom to which R1 and R2 are attached,
in which optionally up to two carbon ring members are replaced by oxygen and/or sulphur and
which is optionally substituted up to three times by identical or different substituents selected from the group consisting of (C1-C6)-alkyl, hydroxyl, oxo, (C1-C6)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl and halogen, and
R3 represents hydrogen or (C1-C6)-alkoxycarbonyl
and their pharmaceutically acceptable salts, hydrates and prodrugs.
Depending on the substitution pattern, the compounds of the formula (I) according to the invention can exist in stereoisomeric forms which are either like image and mirror image (enantiomers) or are not like image and mirror image (diastereomers). The invention relates both to the enantiomers or diastereomers and to their respective mixtures. The racemic forms, like the diastereomers, can be separated in a known manner into the stereoisomerically uniform components.
Furthermore, certain compounds of the formula (I) can be present in tautomeric forms. This is known to the person skilled in the art, and such compounds are likewise included in the scope of the invention.
Pharmaceutically aceeptable, i.e. physiologically safe, salts can be salts of the compounds according to the invention with inorganic or organic acids. Preference is given to salts with inorganic acids such as, for example, hydrochloric acid, hydrobromic acid, phosphoric acid or sulphuric acid, or to salts with organic carboxylic or sulphonic acids such as, for example, acetic acid, propionic acid, maleic acid, fumaric acid, malic acid, citric acid, tartaric acid, lactic acid, benzoic acid, or methanesulphonic acid, ethanesulphonic acid, benzenesulphonic acid, toluenesulphonic acid or naphthalenedisulphonic acid.
Pharmaceutically acceptable salts which may be mentioned include salts with customary bases, such as, for example, alkali metal salts (for example sodium salts or potassium salts), alkaline earth metal salts (for example calcium salts or magnesium salts) or ammonium salts derived from ammonia or organic amines, such as, for example, diethylamine, triethylamine, ethyldiisopropylamine, procaine, dibenzylamine, N-methylmorpholine, dihydroabietylamine or methylpiperidine.
According to the invention, xe2x80x9chydratesxe2x80x9d are forms of the compounds of the formula (I) above which, in the solid or liquid state, form a molecular compound (solvate) by hydration with water. Examples of hydrates are sesquihydrates, monohydrates, dihydrates and trihydrates. Equally suitable are the hydrates of salts of the compounds according to the invention.
According to the invention, xe2x80x9cprodrugsxe2x80x9d are derivatives of the compounds of the formula (I) above which for their part can be biologically active or inactive, but which, following administration, can be converted under physiological conditions (for example metabolically, solvolytically or otherwise) into the corresponding biologically active form.
Halogen represents fluorine, chlorine, bromine and iodine. Preference is given to chlorine and fluorine.
(C1-C10)-Alkyl represents a straight-chain or branched alkyl radical having 1 to 10 carbon atoms. Examples which may be mentioned are: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-octyl and n-decyl. The corresponding alkyl groups having fewer carbon atoms, such as, for example, (C2-C10)-alkyl, (C1-C6)-alkyl and (C1-C4)-alkyl, are derived analogously from this definition. In general, (C1-C4)-alkyl is preferred.
Also derived from this definition is the meaning of the corresponding component of other more complex substituents, such as, for example, in the case of monoalkylamino, di-acylamino, alkylsulphonyl, alkylthio or alkylene, in which an alkyl radical defined as above is attached via two positions
Mono- or di-(C1-C4)-alkylaminocarbonyl represents an amino group which is attached via a carbonyl group and has one or two identical or different straight-chain or branched alkyl substituents having in each case 1 to 4 carbon atoms. Examples which may be mentioned are: methylaminocarbonyl, ethylaminocarbonyl, n-propylaminocarbonyl, isopropyl-aminocarbonyl, t-butylaminocarbonyl, N,N-dimethylaminocarbonyl, N,N-diethyl-aminocarbonyl, N-ethyl-N-methylamino-carbonyl, N-methyl-N-n-propylaminocarbonyl, N-isopropyl-N-n-propylamino-carbonyl and N-t-butyl-N-methylamino-carbonyl.
(C1-C6)-Alkylthio represents a straight-chain or branched alkylthio radical having 1 to 6 carbon atoms. Preference is given to a straight-chain or branched alkylthio radical having 1 to 4 carbon atoms. Examples which may be mentioned are: methylthio, ethylthio, n-propylthio, isopropylthio, t-butylthio, n-pentylthio and n-hexylthio.
(C3-C8)-Cycloalkyl represents a cyclic alkyl radical having 3 to 8 carbon atoms. Examples which may be mentioned are: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. The corresponding cycloalkyl groups having fewer carbon atoms, such as, for example, (C3-C6)-cycloalkyl, are derived analogously from this definition. Preference is given to cyclopropyl, cyclopentyl and cyclohexyl.
The meaning of the corresponding component of other more complex substitutents, such as, for example, in the case of cycloalkylene, in which a cycloalkyl radical as defined above is attached via two positions, is also derived from this definition.
(C1-C6)-Alkoxy represents a straight-chain or branched alkoxy radical having 1 to 6 carbon atoms. Examples which may be mentioned are: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, n-pentoxy and n-hexoxy. The corresponding alkoxy groups having fewer carbon atoms, such as, for example, (C1-C4)-alkoxy, are derived analogously from this definition. In general, (C1-C4)-alkoxy is preferred.
The meaning of the corresponding component of other more complex substitutents, such as, for example, alkoxycarbonyl, in which an alkoxy radical as defined above is attached via a carbonyl group is also derived from this definition.
(C1-C6)-Alkanoyl represents a straight-chain or branched alkyl radical having 1 to 6 carbon atoms which carries a doubly attached oxygen atom in the 1-position and is attached via the 1-position. Examples which may be mentioned are: formyl, acetyl, propionyl, n-butyryl, i-butyryl, pivaloyl and n-hexanoyl. The corresponding alkanoyl groups having fewer carbon atoms, such as, for example, (C1-C4)-alkanoyl, are derived analogously from this definition. In general, preference is given to (C1-C4)-alkanoyl.
(C1-C6)-Alkanoylamido represents an alkanoyl radical as defined above which is attached via an xe2x80x94NHxe2x80x94 group.
(C6-C10-Aryl represents an aromatic radical having 6 to 10 carbon atoms. Examples which may be mentioned are: phenyl and naphthyl.
The meaning of the corresponding component of other more complex substituents, such as, for example, arylene, in which an aryl radical as defined above is attached via two positions, is also derived from this definition.
5- to 13-membered heteroaryl or a 5- to 13-membered aromatic heterocycle having up to 4 heteroatoms from the group consisting of N, O and S represents a mono-, bi- or tricyclic heteroaromatic which is attached via a ring carbon atom of the heteroaromatic or, if appropriate, via a ring nitrogen atom of the heteroaromatic. Examples which may be mentioned are: pyridyl, pyridyl N-oxide, pyrimidyl, pyridazinyl, pyrazinyl, thienyl, furyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl or isoxazolyl, indolicenyl, indolyl, benzo[b]thienyl, benzo[b]furyl, benzothiadiazolyl, indazolyl, quinolyl, isoquinolyl, naphthyridinyl, quinazolinyl. The corresponding heterocycles having a smaller ring size, such as, for example, 5- or 6-membered aromatic heterocycles, or else heterocycles having fewer heteroatoms, such as, for example, those having up to 3 heteroatoms from the group consisting of N, O and S, are derived analogously from this definition. In general, 5- or 6-membered aromatic heterocycles having up to 3 heteroatoms from the group consisting of N, O and S, such as, for example, pyridyl, pyridyl-N-oxide, pyrimidyl, pyridazinyl, furyl, imidazolyl and thienyl are preferred.
The meaning of the corresponding component of other more complex substituents, such as, for example, heteroarylene, in which a heteroaryl radical as defined above is attached via two positions, is also derived from this definition.
A 5- to 13-membered saturated or partially unsaturated heterocycle having up to 4 heteroatoms from the group consisting of N, O and S represents a mono-, bi- or tricyclic heterocycle which may contain one or more double bonds and which is attached via a ring carbon atom or a ring nitrogen atom. Examples which may be mentioned are: tetrahydrofuryl, pyrrolidinyl, pyrrolinyl, piperidinyl, 1,2-dihydropyridinyl, 1,4-dihydropyridinyl, piperazinyl, morpholinyl, morpholinyl-N-oxide, thiomorpholinyl, azepinyl, 1,4-diazepinyl and cyclohexyl. Preference is given to piperidinyl, morpholinyl and pyrrolidinyl.
The corresponding heterocycles having a different ring size, such as, for example, 3- to 7-membered heterocycles, are derived analogously from this definition.
Preference is given to compounds of the formula (I) according to the invention in which
A represents a ring member xe2x80x94CH2xe2x80x94Dxe2x80x94 or xe2x80x94Dxe2x80x94CH2xe2x80x94, in which
D represents xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94,
X represents (C2-C4)-alkylene or cyclohexylene,
R1 represents hydrogen, (C1-C4)-alkyl which is optionally substituted up to three times by fluorine, or (C3-C6)-cycloalkyl,
R2 represents a group of the formula xe2x80x94SO2xe2x80x94R4, xe2x80x94COxe2x80x94R7 or xe2x80x94COxe2x80x94OR10, in which
R4 represents (C1-C4)-alkyl which is optionally substituted up to three times by fluorine, or represents a group of the formula
xe2x80x94Gxe2x80x94E
xe2x80x83in which
E represents (C6-C10)-aryl or a 5- to 13-membered saturated, partially unsaturated or aromatic heterocycle having up to three heteroatoms from the group consisting of N, O and S, where the ring systems are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, fluorine, chlorine, bromine, optionally benzamido-substituted (C1-C4)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, oxo, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl, (C1-C4)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C4)-alkylaminocarbonyl, (C1-C4)-alkanoylamido, (C1-C4)-alkylsulphonyl, (C1-C4)-alkylthio and optionally methyl-, fluorine-, chlorine-, bromine- or nitro-substituted phenylsulphonyl, and
G is absent or represents (C6-C10)-arylene or 5- or 6-membered heteroarylene having up to three heteroatoms from the group consisting of N, O and S, which are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, fluorine, chlorine, bromine, (C1-C4)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl, (C1-C4)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C4)-alkylaminocarbonyl, (C1-C4)-alkanoylamido, (C1-C4)-alkylsulphonyl and (C1-C4)-alkylthio,
R7 represents (C6-C10)-aryl which is optionally substituted by nitro,
R10 represents (C1-C4)-alkyl which is optionally substituted up to three times by chlorine, or
R1 and R2 together with the nitrogen atom to which they are attached form a saturated 5- to 10-membered mono- or bicyclic heterocycle
which has a carbonyl group directly adjacent to the nitrogen atom to which R1 and R2 are attached,
in which optionally up to two carbon ring members are replaced by oxygen and/or sulphur and
which is optionally substituted up to three times by identical or different substituents selected from the group consisting of (C1-C4)-alkyl, hydroxyl, oxo, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl and halogen, and
R3 represents hydrogen
and their pharmaceutically acceptable salts, hydrates and prodrugs.
Particular preference is given to compounds of the formula (I) according to the invention in which
A represents a ring member xe2x80x94CH2xe2x80x94Dxe2x80x94 or xe2x80x94Dxe2x80x94CH2xe2x80x94, in which
D represents xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94 or xe2x80x94Sxe2x80x94,
X represents (C2-C4)-alkylene,
R1 represents hydrogen, (C1-C4)-alkyl which is optionally substituted up to three times by fluorine, or (C3-C6)-cycloalkyl,
R2 represents a group of the formula 
xe2x80x83in which
E represents (C6-C10)-aryl or a 5- to 13-membered saturated, partially unsaturated or aromatic heterocycle having up to three heteroatoms from the group consisting of N, O and S, where the ring systems are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, fluorine, chlorine, bromine, optionally benzamido-substituted (C1-C4)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, oxo, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl, (C1-C4)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C4)-alkylaminocarbonyl, (C1-C4)-alkanoylamido, (C1-C4)-alkylsulphonyl, (C1-C4)-alkylthio and optionally methyl-, fluorine-, chlorine-, bromine- or nitro-substituted phenylsulphonyl, and
G is absent or represents (C6-C10)-arylene or 5- or 6-membered heteroarylene having up to three heteroatoms from the group consisting of N, O and S, which are in each case optionally substituted up to three times by identical or different substituents selected from the group consisting of nitro, cyano, fluorine, chlorine, bromine, (C1-C4)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl, (C1-C4)-alkanoyl, amino, aminocarbonyl, mono- and di-(C1-C4)-alkylaminocarbonyl, (C1-C4)-alkanoylamido, (C1-C4)-alkylsulphonyl and (C1-C4)-alkylthio, or
R1 and R2 together with the nitrogen atom to which they are attached form a saturated 5- to 10-membered mono- or bicyclic heterocycle
which has a carbonyl group directly adjacent to the nitrogen atom to which R1 and R2 are attached,
in which optionally up to two carbon ring members are replaced by oxygen and/or sulphur and
which is optionally substituted up to three times by identical or different substituents selected from the group consisting of (C1-C4)-alkyl, hydroxyl, oxo, (C1-C4)-alkoxy, carboxyl, (C1-C4)-alkoxycarbonyl and halogen, and
R3 represents hydrogen
and their pharmaceutically acceptable salts, hydrates and prodrugs.
Very particular preference is given to compounds of the formula (I) according to the invention in which
A represents a ring member xe2x80x94CH2xe2x80x94Dxe2x80x94 or xe2x80x94Dxe2x80x94CH2xe2x80x94, in which
D represents xe2x80x94CH2xe2x80x94 or xe2x80x94Sxe2x80x94,
X represents (C2-C4)-alkylene,
R1 represents hydrogen, methyl, trifluoromethyl or cyclopropyl.
R2 represents a group of the formula 
xe2x80x83in which
E represents (C6-C10)-aryl or a 5- to 10-membered partially unsaturated or aromatic heterocycle having up to three heteroatoms from the group consisting of N, O and S, where the ring systems are in each case optionally substituted up to two times by identical or different substituents selected from the group consisting of nitro, cyano, amino, fluorine, chlorine, bromine, (C1-C4)-alkyl, trifluoromethyl, (C3-C6)-cycloalkyl, hydroxyl, oxo and (C1-C4)-alkoxy, and
G is absent or represents phenylene or thienylene, and
R3 represents hydrogen,
and their pharmaceutically acceptable salts, hydrates and prodrugs.
Most preference is given to the compounds of Examples, 6, 57, 59, 69, 71, 72 and their pharmaceutically acceptable salts, hydrates and prodrugs.
The present invention also provides a process for preparing the compounds of the formula (I) according to the invention, where
Compounds of the formula (II) 
in which
A is as defined above are reacted with compounds of the formula (III) 
xe2x80x83in which
Y represents R2 or a customary amino protective group and X, R1 and R2 are each as defined above to give compounds of the formula (IV) 
xe2x80x83in which
Y represents R2 or a customary amino protective group and A, X, R1 and R2 are each as defined above
are subsequently reacted with chlorocarbonyl isocyanate to give compounds of the formula (V) 
xe2x80x83in which
Y represents R2 or a customary amino protective group and A, X, R1 and R2 are each defined above,
Y represents a customary amino protective group, compounds of the formula (V) are, if appropriate, converted, by removal of this protective group, into compounds of the formula (VI) 
xe2x80x83in which
A, X and R1 are each as defined above
and are reacted, if appropriate in the presence of a base, with compounds of the formula (VII)
R2xe2x80x94Txe2x80x83xe2x80x83(VII)
xe2x80x83in which
R2 is as defined above and T represents a leaving group to give compounds of the formula (V) in which
Y represents R2 and A, X, R2 and R2 are each as defined above
and, if appropriate, compounds of the formula (V) are reacted, if appropriate in the presence of a base, with compounds of the formula (VIII)
R3xe2x80x94Txe2x80x83xe2x80x83(VIII)
xe2x80x83in which
R3 is as defined above, but not hydrogen, and T represents a leaving group to give compounds of the formula (I) in which R3 is as defined above, but not hydrogen,
where, if the compound prepared in this manner contains a bromoaryl or bromohetaryl group in the radical R3, a transition-metal-catalysed coupling reaction with an organotin or organoboron compound by customary methods may follow and/or
where, if the compound prepared in this manner contains an aldehyde group in the radical R3, this aldehyde group may then be converted into the corresponding oxime using customary methods and/or
where, if the compound prepared in this manner contains a cyano group in the radical R3, this cyano group may then be converted via the stage of the corresponding hydroxylamidine, into the corresponding amidine, using customary methods.
The process according to the invention for preparing compounds of the formula (I) can be illustrated in an exemplary, but not limiting, manner by the equations below: 
Solvents suitable for the process described above are organic solvents which are inert under the reaction conditions. These include halogenated hydrocarbons, such as dichloromethane, trichloromethane, carbon tetrachloride, 1,2-dichloroethane, trichloroethane, tetrachloroethane, 1,2-dichloroethylene or trichloroethylene, ethers, such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether or diethylene glycol dimethyl ether, hydrocarbons, such as benzene, xylene, toluene, hexane or cyclohexane, dimethylformamide, dimethyl sulphoxide, acetonitrile, pyridine or hexamethylphosphoric triamide. It is also possible to use solvent mixtures of the solvents mentioned above.
The reactions are generally carried out in a temperature range of from xe2x88x9278xc2x0 C. to reflux temperature, preferably in the range from 0xc2x0 C. to reflux temperature.
The reactions can be carried out at atmospheric, elevated or reduced pressure (for example in the range from 0.5 to 5 bar). In general, they are carried out at atmospheric pressure.
Suitable bases are the customary inorganic or organic bases. These preferably include alkali metal hydroxides, such as, for example, sodium hydroxide or potassium hydroxide, or alkali metal carbonates, such as sodium carbonate or potassium carbonate, or sodium methoxide or potassium methoxide or sodium ethoxide or potassium ethoxide or potassium tert-butoxide, or amides, such as sodium amide, lithium bis-(trimethylsilyl)amide or lithium diisopropylamide, or amines, such as triethylamine, diisopropylethylamine, diisopropylamine, 4,N,N-dimethylaminopyridine or pyridine.
Preferred solvent for the reaction of compounds of the formula (II) with compounds of the formula (III) to give compounds of the formula (IV) and the further conversion with chlorocarbonyl isocyanate and to compounds of the formula (V) is toluene. In the reaction of compounds of the formula (VI) with compounds of the formula (VII), pyridine is preferred if R2 is a sulphonyl radical and tetrahydrofuran, dichloromethane or acetonitrile is preferred if R2 is an alkoxycarbonyl radical.
The temperature range for the reaction of compounds of the formula (II) with compounds of the formula (III) to give compounds of the formula (IV) is in particular from 80 to 120xc2x0 C. The addition of chlorocarbonyl isocyanate in the conversion of compounds of the formula (IV) into compounds of the formula (V) is in particular carried out at room temperature; the further reaction is then carried out in particular in a temperature range from 80 to 120xc2x0 C. The reaction of compounds of the formula (VI) with compounds of the formula (VII) is, if R2 is a sulphonyl radical, carried out in particular at room temperature.
The reaction of compounds of the formula (II) with compounds of the formula (III) to give compounds of the formula (IV) may, if appropriate, be accelerated by adding catalytic amounts of acid, preferably organic sulphonic acid, in particular camphorsulphonic acid.
Suitable leaving groups T in compounds of the formulae (VII) and (VIII) are, for example: halogen, mesylate, tosylate, triflate or 1-imidazolyl, preferably chlorine.
Suitable amino protective groups for compounds of the formulae (III), (IV) and (V) are the radicals which are customarily used, as described, for example, in T. W. Greene, P. G. M. Wuts, Protective Groups in Organic Synthesis, second edition, New York, 1991. Examples which may be mentioned are: benzyl and BOC (tert-butoxycarbonyl). Removal of the amino protective group in the conversion of compounds of the formula (V) into compounds of the formula (VI) is carried out in the manner customary for the amino protective group in question, as described, for example, in T. W. Greene, P. G. M. Wuts, Protective Groups in Organic Synthesis, second edition, New York, 1991.
The transition-metal-catalysed coupling reaction, which is carried out, if appropriate, of aryl or hetaryl bromides with organotin compounds (Stille coupling) or organoboron compounds (Suzuki coupling) is carried out under the reaction conditions customary for these couplings, in the presence of a catalyst, preferably in the presence of a transition metal catalyst, in particular in the presence of a palladium catalyst (see, for example J. Tsuju, Palladium Reagents and Catalysts, J. Wiley and Sons, 1995), preferably in the solvent dimethylformamide. Preferred transition metal catalysts are palladium(0) or palladium(II) compounds, in particular bis-(triphenylphosphine)-palladium(II) chloride or tetrakis-(triphenylphosphine)-palladium(0). If organoboron compounds are used (Suzuki coupling, review: N. Miyaura, A. Suzuki, Chem. Rev. 1995, 95, 2457-2483), the reaction is carried out, in particular, at a temperature of from 90xc2x0 C. to 110xc2x0 C. in the presence of a base, preferably aqueous sodium carbonate solution. If organotin compounds are used (Stille coupling, review: V. Farina, V. Krishnamurthy, W. J. Scott in: The Stille Reaction, 1998, J. Wiley and Sons, New York), the reaction is carried out, in particular, at a temperature of from 110 to 130xc2x0 C.
The conversion of aldehyde groups into the corresponding oximes and of cyano groups, via the stage of the corresponding hydroxyamidines, into the corresponding amidines, are carried out, if appropriate, using the preparation methods customary for these reactions. Furthermore, reference is made to Examples 108 to 110 in the experimental part.
The compounds of the formulae (II), (III), (VII) and (VIII) are known to the person skilled in the art or can be prepared by customary methods.
The compounds of the formula (I) according to the invention have an unforeseeable useful spectrum of pharmacological activity, and they are therefore particularly suitable for the prophylaxis and/or treatment of disorders.
They can preferably be used in medicaments for the prevention and/or therapy of ischaemic and reperfusion damage in the heart (after an acute infarction), in the brain (after a stroke) or in skeletal muscle, for cardiovascular disorders, such as, for example, unstable angina pectoris and arteriosclerosis, neuronal and neurodegenerative disorders, such as, for example, epilepsy, chronic pain, Alzheimer""s disease and Parkinson""s disease, traumatic brain injuries, septic shock, and also arthritis, diabetes, chronic colitis, sudden deafness, inflammable pulmonary disorders, such as, for example, asthma and chronic bronchitis, and cancer.
The present invention also relates to the use of the substances of the formula (I) for preparing medicaments and to pharmaceutical compositions for the prophylaxis and/or treatment of the clinical pictures mentioned above.
The present invention furthermore relates to a method for the prophylaxis and/or treatment of the clinical pictures mentioned above using the substances of the formula (I).
In addition, the compounds according to the invention can also be used for the treatment of acute myocardial infarction, including in combination with one or more of the following medicaments which are used for the standard therapy of acute myocardial infarction: calcium channel blockers (such as, for example, nifedipine, diltiazem, verapamil), nitrovasodilators (such as, for example, isosorbide dinitrate, glycerol trinitrate, isosorbide 5-mono-nitrate, molsidomine), beta blockers, (such as, for example, metoprolol, atenolol, propranolol, solatol), platelet aggregation inhibitors (such as, for example, acetylsalicylic acid, triclopidine, clopidrogrel), thrombolytics (fibrinolytics) (such as, for example, streptokinase, alteplase, reteplase, urokinase, anistreplase), anticoagulants (such as, for example, heparin, warfarin, phenprocoumarin, low-molecular-weight heparins), ACE inhibitors (such as, for example, enalapril), glycoprotein IIb/IIIa receptor antagonists (such as, for example, tirofiban, eptifibatide), antiarrhythmics (such as, for example, lidocaine, amiodarone) and beta-adrenergic agonists (such as, for example, dopamine, dobutamine).
1) Test Description PARS Inhibition Test (in vitro)
The activity of substances as PARS inhibitors is tested in accordance with the method of Ushiro [Ushiro et al., J. Biol. Chem., 262, 2352-2357 (1987)]. To this end, recombinantly expressed (Bac-To-Bac, Baculo virus expression system; Instruction Manual; Life Technologies) human PARS enzyme is activated in a buffer which contains radioactively labelled [14C]-NAD+. The poly(ADP-ribose) units that are synthesized are precipitated using trichloroacetic acid, and the proportion of labelled protein is determined by scintillation measurements. Incubation of PARS with inhibitors leads to a decrease in the proportion of labelled protein and thus to a reduced radioactivity.
Inhibition of the PARS activity can be represented as a percentage of PARS inhibition in incubation with different substances or as the concentration at which 50% of the enzyme is inhibited, i.e. as IC50 value.
Material
Buffer:
100 mM
Tris-HCl,
pH 7.4
10 mM
MgCl2 
1 mM
dithiothreitol (DTT)
Tris-HCl and MgCl2 are dissolved in water, DTT is added from an aqueous 100 mM stock solution (stored at xe2x88x9220xc2x0 C.) and the pH is adjusted with concentrated HCl to 7.4.
DNA:
1 mg/ml of calf thymus DNA
1 mg/ml of calf thymus DNA (from Sigma) is dissolved in water and sonicated to induce strand breaks. 500 xcexcl aliquots were stored at xe2x88x9220xc2x0 C.
Histones:
10 mg/ml of type IIA histones, calf thymus
10 mg/ml of lyophilized histones (from Sigma) are dissolved in water.
500 xcexcl aliquots are stored at xe2x88x9220xc2x0 C.
NAD+ Mix:
2 mM NAD+ in buffer,
NAD+ (from Sigma) solutions are prepared freshly before each test.
In each case 3 xcexcl of labelled [14C]-NAD+ (2.8 kBq, from Amersham) are added to 7 xcexcl of cold NAD+ solution.
Trichloroacetic acid (TCA):
TCA is stored at 4xc2x0 C. as a 10% strength by weight solution.
PARS:
Human PARS protein is expressed recombinantly in the baculo virus system (Bac-To-Bac, Baculo virus expression system; Instruction Manual; Life Technologies) and purified. 500 xcexcl aliquots are stored at xe2x88x9280xc2x0 C.
Methods
The compounds to be tested are dissolved in DMSO (dimethyl sulphoxide) at a concentration of 10 mM. The assay is carried out in deep 96-well plates. Per well, 70 xcexcl of buffer, 10 xcexcl of DNA, 10 xcexcl of histones, 10 xcexcl of NAD+/[14C]-NAD+ mix and 0.5-5 xcexcl of PARS (about 10,000 cpm/test) are combined with 1 xcexcl of the compounds (final concentration 0.001-10 xcexcM), to give a total volume of about 110 xcexcl. The mixture is incubated at room temperature for 10 min, and 1 xcexcl of ice-cold TCA solution is then added, and the precipitated labelled proteins are sucked onto a filter paper (printed filter mat A; from Wallac) using a harvester (from Scatron). The filter is dried, sealed together with a scintillation sheet (Multilex A; from Wallac) and measured in a xcex2 counter for 1 min per well.
Results of the PARS Inhibition Test
In addition to the substances described in this application, the known PARS inhibitor 1,5-dihydroxyisoquinoline (DHCH) is tested as reference substance. The results of the test are stated as IC50 values for the inhibition of PARS.
The results are shown in Table 1:
2) Test Description Cell Protection Assay (in vitro)
In accordance with a method described by Bowes [Bowes et al., Br. J. Pharmacol., 124, 1760-1766 (1998)], the ability of PARS inhibitors to protect cells against cell death induced by incubation with H2O2 is examined in a cell protection assay. Incubation of endothelial cells with H2O2 results in the generation of DNA strand breaks which in turn activate PARS, resulting in a drastic energy decrease in the cells and in cell death. Living cells are quantified by a fluorimetric redox indicator (Alamar blue), which is converted in the electron transport system of the mitochondria.
Specifically, 7500 MHEC5-T cells/well (DSM ACC 336; German collection of microorganisms and cell cultures) are sown in 4 replications on a 96-well plate. After 24 hours, the cells are incubated with 3 mM aqueous H2O2 solution and differing concentrations of the substances in the presence of 6% by volume of Alamar blue solution in the medium at 37xc2x0 C. for 5 hours. The reference substance used is 10 xcexcM 1,5-dihydroxyisoquinoline (DHCH) solution. After the incubation, the fluorescence is measured at an excitation wavelength of 530-560 nm and an emission wavelength of 590 nm. The percentage for the cell protection is calculated as the difference between the living cells treated only with H2O2 and the cells treated with H2O2 and PARS inhibitors. The internal standard used is 10 xcexcM DHCH, which is defined as 100% protection. The results obtained for the other substances are compared to this value.
Results of the cell protection assays:
Examples of the protection of endothelial cells by PARS inhibitors are listed in Table 2 below. The EC50 values indicate the concentration at which 50% of maximum cell protection is reached, maximum protection by 10 xcexcM DHCH being defined as 100%. DHCH has an EC50 value of 2 xcexcM.
3) Test Description xe2x80x9cWorking Heartxe2x80x9d Model (in vivo)
For tests on isolated hearts in the xe2x80x9cworking heartxe2x80x9d mode [Bardenheuer and Schrader, Circulation Res., 51, 263 (1983)], isolated hearts of rats are subjected to a 60-minute xe2x80x9clow-flowxe2x80x9d phase to generate global ischaemia, and the action of the substances with respect to the reestablishment of the pressure in the left ventricle (LVPmax) and the contractile force (dP/dt) during the reperfusion phase is examined. The control substance used is 1,5-dihydroxyisoquinoline.
The present invention also relates to medicaments and pharmaceutical compositions comprising at least one compound of the formula (I), preferably together with one or more pharmacologically acceptable auxiliaries or excipients, and to their use for the abovementioned purposes.
The active compound can act systemically and/or locally. To this end, it can be administered in a suitable manner, such as, for example, orally, parenterally, pulmonarily, nasally, sublingually, lingually, buccally, rectally, transdermally, conjunctivally, otically or as an implant.
For these administration routes, the active compound can be administered in suitable administration forms.
Administration forms suitable for oral administration are known administration forms which release the active compound rapidly and/or in modified form, such as, for example, tablets (uncoated and also coated tablets, for example enterically coated tablets), capsules, sugar-coated tablets, granules, pellets, powders, emulsions, suspensions and solutions.
Parenteral administration can be effected by circumventing a bioabsorption step (in an intravenous, intraarterial, intracardial, intraspinal or intralumbal manner), or via bioabsorption (intramuscularly, subcutaneously, intracutaneously, percutaneously or intraperitoneally). Administration forms suitable for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophilisates and sterile powders.
Medicinal forms suitable for the other administration routes are, for example, medicinal forms for inhalation (inter alia powder inhalators, nebulizers), nasal drops/solutions, sprays; tablets or capsules to be administered lingually, sublingually or buccally, suppositories, preparations for ears or eyes, vaginal capsules, aqueous suspensions (lotions, agitated mixtures), lipophilic suspensions, ointments, creams, milk, pastes, powder for spreading or implants.
The active compounds can be converted in a manner known per se into the administration forms listed. This is effected using inert non-toxic, pharmaceutically suitable auxiliaries. These include, inter alia, excipients (for example microcrystalline cellulose), solvents (for example liquid polyethylene glycols), emulsifiers (for example sodium dodecyl sulphate), dispersants (for example polyvinylpyrrolidone), synthetic and natural biopolymers (for example albumin), stabilizers (for example antioxidants such as ascorbic acid), colorants (for example inorganic pigments, such as iron oxides), or flavour- and/or odour-masking substances.
In the pharmaceutical preparations listed above, the therapeutically active compounds should be present in a concentration of from about 0.1 to 99.5, preferably from about 0.5 to 95, % by weight of the total mixture, i.e. the active compound should be present in amounts sufficient to achieve the dosage range indicated.
In addition to the compounds of the formula (I) according to the invention, the pharmaceutical preparations listed above may also comprise other pharmaceutically active compounds.
In general, it has been found to be advantageous both in human and veterinary medicine to administer the active compound(s) according to the invention in total amounts of from about 0.1 to about 500, preferably from 1 to 100, mg/kg of body weight per 24 hours, if appropriate in the form of a plurality of individual doses, to obtain the desired results. An individual dose preferably comprises the active compound(s) according to the invention in amounts of from about 0.1 to 80, in particular from 1 to 30, mg/kg of body weight.
In spite of this, it may be necessary, if appropriate, to depart from the amounts mentioned, namely depending on the body weight or on the type of administration route, on the individual response to the medicament, the manner of its formulation and the time or interval at which administration takes place. Thus, in some cases it may be adequate to manage with less than the abovementioned minimum amount, while in other cases the upper limit mentioned has to be exceeded.